The rational design of a Rhodamine fluorescent probe enables the selective detection and bioimaging of salicylic acid in plants under abiotic stress

Abiotic stress severely hinders plant growth and development, resulting in a considerable reduction in crop yields. Salicylic acid (SA) serves as a central signal mediating abiotic stress responses in plants. Real-time fluorescence tracking using specific probes can enhance our understanding of the SA-triggered modulation underlying these events. However, in complicated living plant microenvironments, selective recognition and bioimaging of SA is a great challenge for scientists due to the severe background interference and SA analogues. Herein, an efficient fluorescence probing technology employing a highly selective rhodamine probe—phoxrodam was developed, which realizes the precise bioimaging of SA in salt-stressed plant seedlings. Experimental findings reveal that phoxrodam demonstrates exceptional selectivity (fluorescence intensity: I_Phoxrodam+SA/I_{Phoxrodam+SA analogues} > 4.29-fold), high sensitivity (limit of detection = 6.42 nM, fluorescence quantum yield: Φ_Phoxrodam+SA = 0.36) and good anti-interference properties. Furthermore, we confirmed that phoxrodam accurately detects SA in the roots of salt-stressed wheat seedlings, the low-temperature resistance of Nicotiana benthamiana and the heavy metal resistance of pea seeds, using in vivo confocal imaging. This study provides a feasible strategy for efficiently tracking plant signalling molecules and promotes the in-depth research of SA-mediated physiological mechanisms, laying a key foundation for the future development of new immune activation inducers.